Trimer stabilizing HIV envelope protein mutations

Abstract

Human immunodeficiency virus (HIV) envelope proteins having specified mutations that stabilize the trimeric form of the envelope protein are provided. The HIV envelope proteins described herein have an improved percentage of trimer formation and/or an improved trimer yield. Also provided are particles displaying the HIV envelope proteins, nucleic acid molecules and vectors encoding the HIV envelope proteins, as well as compositions containing the HIV envelope proteins, particles, nucleic acid, or vectors.

It is claimed: 
     
       1. A trimeric complex comprising a noncovalent oligomer of three recombinant human immunodeficiency virus 1 (HIV-1) envelope (Env) proteins, wherein the amino acid at position 658 of each of the HIV-1 Env proteins is mutated to an amino acid selected from the group consisting of Val, Ile, Phe, Met, and Ala; wherein the numbering of the positions is according to the numbering in gp160 of HIV-1 isolate HXB2 as shown in SEQ ID NO: 1. 
     
     
       2. The trimeric complex of  claim 1 , wherein the amino acid at position 658 is Val. 
     
     
       3. The trimeric complex of  claim 1 , wherein the amino acid at position 658 is Ile. 
     
     
       4. The trimeric complex of  claim 1 , wherein each of the HIV-1 Env proteins is a clade A, B, or C HIV-1 Env protein. 
     
     
       5. The trimeric complex of  claim 1 , wherein each of the HIV-1 Env proteins further comprises Cys at positions 501 and 605 or Pro at position 559, or Cys at positions 501 and 605 and Pro at position 559. 
     
     
       6. The trimeric complex of  claim 1 , wherein each of the HIV-1 Env proteins further comprises a replacement of the furin cleavage sequence at positions 508-511 by RRRRRR (SEQ ID NO: 10). 
     
     
       7. The trimeric complex of  claim 1 , wherein each of the recombinant HIV-1 Env proteins is a gp140 or gp160 protein. 
     
     
       8. A composition comprising the trimeric complex of  claim 1 , and a pharmaceutically acceptable carrier. 
     
     
       9. A liposome or nanoparticle displaying on its surface the trimeric complex of  claim 1 . 
     
     
       10. A trimeric complex comprising a noncovalent oligomer of three identical recombinant human immunodeficiency virus 1 (HIV-1) envelope (Env) proteins, wherein the amino acid at position 658 of each of the HIV-1 Env proteins is mutated to an amino acid selected from the group consisting of Val, Ile, Phe, Met, Ala, and Leu, and each of the recombinant HIV-1 Env proteins further comprises one or more of the following amino acid residues:
 (i) Phe, Leu, Met, or Trp, at position 651; 
 (ii) Phe, Ile, Met, or Trp, at position 655; 
 (iii) Asn or Gln, at position 535; 
 (iv) Val, Ile or Ala at position 589; 
 (v) Phe or Trp, at position 573; 
 (vi) Ile at position 204; 
 (vii) Phe, Met, or Ile, at position 647; 
 (viii) Gln, Glu, Ile, Met, Val, Trp, or Phe, at position 588; 
 (ix) Lys at position 64 or Arg at position 66 or Lys at position 64 and Arg at position 66; 
 (x) Trp at position 316; 
 (xi) Cys at both positions 201 and 433; 
 (xii) Pro at position 556 or 558 or at both positions 556 and 558; 
 (xiii) replacement of the loop at amino acid positions 548-568 (HR1-loop) by a loop having an amino acid sequence selected form the group consisting of SEQ ID NOs 12-17; 
 (xiv) Gly at position 568, or Gly at position 569, or Gly at position 636, or Gly at both positions 568 and 636, or Gly at both positions 569 and 636; 
 (xv) Tyr at position 302, or Arg at position 519, or Arg at position 520, or Tyr at position 302 and Arg at position 519, or Tyr at position 302 and Arg at position 520, or Tyr at position 302 and Arg at both positions 519 and 520; and/or 
 (xvi) Cys at positions 501 and 605, or Pro at position 559, or Cys at positions 501 and 605 and Pro at position 559, 
 
       wherein the numbering of the positions is according to the numbering in gp160 of HIV-1 isolate HXB2 as shown in SEQ ID NO: 1. 
     
     
       11. The trimeric complex of  claim 10 , wherein each of the recombinant HIV-1 Env proteins comprises Val at position 658 and Ile at position 655. 
     
     
       12. The trimeric complex of  claim 10 , wherein each of the recombinant HIV-1 Env proteins comprises Val at position 658 and Phe at position 651. 
     
     
       13. The trimeric complex of  claim 10 , wherein each of the recombinant HIV-1 Env proteins comprises at least two of the amino acid residues of (i) to (vii). 
     
     
       14. The trimeric complex of  claim 10 , wherein each of the recombinant HIV-1 Env proteins comprises Cys at positions 501 and 605, or Pro at position 559, or Cys at positions 501 and 605 and Pro at position 559. 
     
     
       15. A method of improving the trimer formation of an human immunodeficiency virus 1 (HIV-1) envelope (Env) protein, the method comprising introducing a substitution of Lys or Gln at position 658 of the HIV-1 Env protein by Val, Ile, Phe, Met, Ala, or Leu, into the HIV-1 Env protein, wherein the numbering of the position is according to the numbering in gp160 of HIV-1 isolate HXB2 as shown in SEQ ID NO: 1.